**4. Phytosanitary diagnosis**

Diseases represent one of the main factors that affect sugarcane production. Knowledge of phytosanitary status and the correct identification of phytopathogens are key to reducing losses due to diseases. In this regard, it is essential to carry out a phytosanitary diagnosis at an early stage to ensure the phytosanitary quality of the seedlings obtained in the laboratory.

In many countries where sugarcane is an important part of the economy, plant health departments (PHDs) are responsible for certifying the procedures for obtaining pest- and diseasefree sugarcane vitroplants. The PHD determines the requirements necessary for accreditation of micropropagation laboratories engaged in in vitro culture of sugarcane, whose legal basis is determined based on Plant Health Laws. To obtain certification, a laboratory must meet a series of requirements that demonstrate technical competence, satisfactory infrastructure and sanitary capacity to produce plant material in vitro.

Accreditation of micropropagation laboratory: micropropagation laboratory should be accredited by an appropriate authority to ensure technical competence and satisfactory infrastructure.


To determine plant health, samples of sugarcane shoots are taken *in vitro* and sent to the Phytosanitary Diagnostic Centers authorized to verify the plant health. *In vitro* propagated plants should be indexed for freedom from viruses and virus-like diseases through enzyme-

*In Vitro* Propagation of Sugarcane for Certified Seed Production

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**Figure 3.** Certified vitroplants obtained from laboratory. (a) Vitroplants ready for field transfer and (b) field planting.

According to the phytosanitary diagnosis report and based on the Plant Health Law, the diag-

After obtaining healthy in vitro plants, a certification is issued so that vitroplants can be used

Seed production system comprises basic seed (primary seed), semicommercial seed (founda-

The unit area of the seed nursery should be approximately one-tenth of the area that is planned to be renewed each year in the commercial plantations. This plantation system is used because

linked immunosorbent assay (ELISA) and molecular methods [23].

nosis must be negative for the main sugarcane diseases (**Table 2**).

for the establishment of certified basic seed nurseries (**Figure 3**).

tion or secondary seed) and commercial seed production.

**5. Seed production**

**Table 2.** Pests and diseases which plant material produced *in vitro* must be free of.

**Figure 3.** Certified vitroplants obtained from laboratory. (a) Vitroplants ready for field transfer and (b) field planting.

To determine plant health, samples of sugarcane shoots are taken *in vitro* and sent to the Phytosanitary Diagnostic Centers authorized to verify the plant health. *In vitro* propagated plants should be indexed for freedom from viruses and virus-like diseases through enzymelinked immunosorbent assay (ELISA) and molecular methods [23].

According to the phytosanitary diagnosis report and based on the Plant Health Law, the diagnosis must be negative for the main sugarcane diseases (**Table 2**).

After obtaining healthy in vitro plants, a certification is issued so that vitroplants can be used for the establishment of certified basic seed nurseries (**Figure 3**).
